本文選題：血管內(nèi)皮生長因子　切入點：腫瘤轉(zhuǎn)移相關(guān)基因MTA1　出處：《武漢大學》2012年博士論文

【摘要】：第一部分人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA表達的相關(guān)性研究 目的：研究人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與血管內(nèi)皮生長因子(VEGF)、增殖細胞核抗原(PCNA)表達的相關(guān)關(guān)系,并結(jié)合文獻探討其意義。 方法：采用免疫組化(SABC)和圖像分析技術(shù)檢測10例正常鼻咽部組織和75例人鼻咽癌標本中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA表達水平,并分析表達的相關(guān)性。 結(jié)果：(1)腫瘤轉(zhuǎn)移相關(guān)基因MTA1、VEGF、PCNA在正常鼻咽部組織中的表達均為陰性；(2)腫瘤轉(zhuǎn)移相關(guān)基因MTA1、VEGF、PCNA在人鼻咽癌中均有陽性表達,(3)人鼻咽癌中腫瘤轉(zhuǎn)移相關(guān)基因MTA1與VEGF、PCNA的表達水平呈明顯正相關(guān)(r=0.941和r=0.889,P0.05)。 結(jié)論：腫瘤轉(zhuǎn)移相關(guān)基因MTA1在人鼻咽癌的惡性生物學行為中可能起重要的作用,其可能成為人鼻咽癌治療的一個有效的靶點。 第二部分腫瘤轉(zhuǎn)移相關(guān)基因MTA1與鼻咽癌細胞浸潤和轉(zhuǎn)移的關(guān)系 目的：鼻咽癌具有很高的侵潤和轉(zhuǎn)移特性。而腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)是新近發(fā)現(xiàn)的一個腫瘤轉(zhuǎn)移候選基因。本研究通過比較MTA1基因在人鼻咽癌細胞高低轉(zhuǎn)移株的表達水平,探討MTA1表達與鼻咽癌細胞浸潤和轉(zhuǎn)移潛能的相關(guān)性。 方法：采用半定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)檢測增殖及轉(zhuǎn)移潛能不同的鼻咽癌細胞株6-10B和5-8F中MTA1的表達情況,用Boyden小室體外侵襲實驗檢測兩株細胞的體外轉(zhuǎn)移能力；用MTA1基因質(zhì)粒轉(zhuǎn)染低轉(zhuǎn)移潛能細胞株6-10B,通過半定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)檢測MTA1的表達；并檢測轉(zhuǎn)染前后細胞轉(zhuǎn)移能力的變化。 結(jié)果：MTA1在低轉(zhuǎn)移潛能細胞株6-10B中表達水平低,在高轉(zhuǎn)移潛能細胞株5-8F中表達水平高(p0.05)；體外侵襲實驗顯示高轉(zhuǎn)移株5-8F細胞體外侵襲力強(穿膜細胞相對百分率為46.3+2.4%)顯著高于低轉(zhuǎn)移潛能細胞株,其穿膜細胞相對百分率只有12.6+1.1%,轉(zhuǎn)染MTA1基因質(zhì)粒后,低轉(zhuǎn)移細胞株轉(zhuǎn)移潛能較未轉(zhuǎn)染細胞明顯增高(p0.05)。 結(jié)論：MTA1在人鼻咽癌細胞轉(zhuǎn)移過程中起重要作用,其作用機制以及作為腫瘤轉(zhuǎn)移治療靶點的可能性值得進一步研究。 第三部分反義寡核苷酸對鼻咽癌細胞MTAl基因表達及侵襲性影響的研究 目的：合成人腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)的反義脫氧寡核苷酸,觀察其轉(zhuǎn)染后對人鼻咽癌CNE1細胞系中MTA1表達及鼻咽癌侵襲性的影響。 方法：免疫細胞染色觀察人工合成正義、反義及無意義MTA1基因片段轉(zhuǎn)染人鼻咽癌細胞CNE1后人腫瘤轉(zhuǎn)移相關(guān)基因的表達,RT-PCR和Western blot檢測MTA1基因的(?)nRNA和蛋白質(zhì)的表達水平的變化。應(yīng)用Boyden小室體外侵襲力評價轉(zhuǎn)染前后細胞侵襲力的變化。 結(jié)果：反義寡核苷酸處理鼻咽癌細胞后,MTA1mRNA的表達下降(吸光度之比為24.09±0.22),與正義鏈組、無意義鏈組和空白對照組(34.23+0.15,33.22±0.28,35.18±0.22)相比差異有統(tǒng)計學意義,P0.05。Boyden小室體外侵襲實驗檢測顯示,反義寡核苷酸處理后鼻咽癌細胞的透膜能力明顯下降。 結(jié)論：MTA1同鼻咽癌的轉(zhuǎn)移能力密切相關(guān),反義寡核苷酸的轉(zhuǎn)染可阻遏鼻咽癌細胞中人腫瘤轉(zhuǎn)移相關(guān)基因(MTA1)的表達。并因此有可能限制鼻咽癌的侵襲和轉(zhuǎn)移。
[Abstract]:The correlation of tumor metastasis related gene MTA1 and VEGF, PCNA expression in human nasopharyngeal carcinoma
Objective: To investigate the correlation between tumor metastasis related gene MTA1 and vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) expression in human nasopharyngeal carcinoma, and to explore its significance combined with literature.
Methods: immunohistochemistry and SABC were used to detect the expression levels of MTA1 and VEGF and PCNA in 10 normal nasopharyngeal tissues and 75 NPC cases, and the correlation between them was analyzed.
Results: (1) tumor metastasis related genes MTA1, VEGF, PCNA expression in normal nasopharyngeal tissues were negative; (2) tumor metastasis related genes MTA1, VEGF, PCNA were positive in human nasopharyngeal carcinoma, (3) human nasopharyngeal carcinoma metastasis related genes MTA1 and VEGF showed a positive correlation the expression level of PCNA (r=0.941, r=0.889, P0.05).
Conclusion: tumor metastasis related gene MTA1 may play an important role in malignant biological behavior of human nasopharyngeal carcinoma. It may become an effective target for treatment of NPC.
The relationship between the second part of tumor metastasis related gene MTA1 and the invasion and metastasis of nasopharyngeal carcinoma cells
Objective: nasopharyngeal carcinoma with invasion and metastasis is very high. And the characteristics of tumor metastasis associated gene (MTA1) is a recently discovered tumor metastasis genes. The expression level of MTA1 gene in human nasopharyngeal carcinoma cell lines with different metastatic cells, to investigate the correlation between MTA1 expression and the potential of invasion and metastasis of nasopharyngeal carcinoma.
Methods: using semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to detect the proliferation and metastatic potential of different nasopharyngeal carcinoma cell lines 6-10B and MTA1 in 5-8F expression, with Boyden chamber in vitro invasion assay of two cell lines in vitro metastasis; MTA1 gene transfected with low metastatic potential cell line 6-10B was detected by semi quantitative MTA1. Reverse transcriptase polymerase chain reaction; change detection and transfer ability before and after transfection.
Results: the MTA1 in low metastatic potential cell line 6-10B low expression in highly metastatic potential cell line 5-8F high expression level (P0.05); experiments show high metastatic 5-8F cells in vitro invasiveness in vitro invasion (transmembrane cell relative percentage of 46.3+2.4%) was significantly higher than that of low metastatic potential cell line, the cell membrane the relative percentage of only 12.6+1.1%, transfection of MTA1 gene plasmid, metastatic potential than non transfected cells was significantly higher in low metastatic cell line (P0.05).
Conclusion: MTA1 plays an important role in the metastasis of human nasopharyngeal carcinoma cells, and its mechanism and the potential as a therapeutic target for tumor metastasis are worth further research.
Study on the effect of third antisense oligonucleotides on MTAl gene expression and invasiveness in nasopharyngeal carcinoma cells
Objective: to synthesize antisense oligodeoxynucleotide of human tumor metastasis related gene (MTA1) and observe its effect on MTA1 expression and invasion of nasopharyngeal carcinoma cell line after transfection in human nasopharyngeal carcinoma CNE1 cell line.
Methods: To observe the effect of synthetic justice immune cells staining. The expression of antisense MTA1 gene transfection and significance of human nasopharyngeal carcinoma cells CNE1 human tumor metastasis related gene, detection of MTA1 gene RT-PCR and blot Western (?) expression of nRNA and protein. The application of Boyden chamber invasion force evaluation changes before and after transfection cell invasion.
Results: antisense oligonucleotide treatment of nasopharyngeal carcinoma cells, the expression of MTA1mRNA decreased (absorbance ratio of 24.09 + 0.22), and sense group, no significant chain group and blank control group (34.23+0.15,33.22 + 0.28,35.18 + 0.22) compared to the difference was statistically significant, P0.05.Boyden chamber in vitro invasion assay showed that antisense oligonucleotide treatment membrane ability of nasopharyngeal carcinoma cells decreased significantly.
Conclusion: MTA1 is closely related to the metastatic ability of nasopharyngeal carcinoma. The transfection of antisense oligonucleotides can inhibit the expression of human tumor metastasis related gene (MTA1) in nasopharyngeal carcinoma cells. Therefore, it may limit the invasion and metastasis of nasopharyngeal carcinoma.

【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R739.63

【參考文獻】

相關(guān)期刊論文 前10條

1 王會河;黃光武;莫立根;趙惠柳;鄺國乾;;MTA1表達與鼻咽癌浸潤轉(zhuǎn)移的關(guān)系[J];廣西醫(yī)科大學學報;2006年04期

2 王俊平;陳奎生;;食管鱗癌組織中MTA1蛋白的表達[J];醫(yī)藥論壇雜志;2007年01期

3 彭爭榮;鐘衛(wèi)紅;劉娟;肖平田;;高壓氧和5-氟尿嘧啶聯(lián)合作用對鼻咽癌細胞增殖與侵襲遷移的影響[J];臨床耳鼻咽喉頭頸外科雜志;2010年09期

4 曾恒;陳安民;李鋒;楊彩虹;;反義寡核苷酸對骨肉瘤細胞MTA1基因表達及侵襲性影響的研究[J];中華腫瘤防治雜志;2009年04期

5 ;Perspectives on the treatment of colorectal carcinoma[J];World Journal of Gastrointestinal Oncology;2010年05期

6 汪盛;彭程;蔣先鎮(zhèn);陳合群;劉玉明;;腫瘤轉(zhuǎn)移基因MTA1在膀胱癌中的表達及其臨床意義[J];中國醫(yī)學工程;2007年07期

7 李新志;陳安民;易成臘;郭風勁;羅正強;黃仕龍;徐衛(wèi)國;;MTA1基因表達與人骨肉瘤細胞浸潤和轉(zhuǎn)移的關(guān)系[J];中國癌癥雜志;2006年02期

8 林川,陳漢,吳孟超,楊廣順,胡世杰,戴建新;腫瘤轉(zhuǎn)移基因MTA1的分子克隆及其在肝癌組織中表達的初步研究[J];中華普通外科雜志;2000年10期

9 阮洋;王景濤;高艷華;欒海艷;齊淑芳;劉偉新;;Livin、Caspase-3、MTA1在結(jié)腸癌組織中的表達及相關(guān)性的研究[J];中國老年學雜志;2007年13期

10 ;Identification of differential gene expressions in colorectal cancer and polyp by cDNA microarray[J];World Journal of Gastroenterology;2012年06期

，

本文編號：1658943